Differentiation of stem cells into insulin-producing cells under the influence of nanostructural polyoxometalates.

Two polyoxometalates (POMs) with W were synthesized by a two-step, self-assembling method. They were used for stimulation of mesenchymal stem cell differentiation into insulin-producing cells. The nanocompounds (tris(vanadyl)-substituted tungsto-antimonate(III) anions [POM1] and tris-butyltin-21-tungsto-9-antimonate(III) anions [POM2]) were characterized by analytical techniques, including ultraviolet-visible, Fourier transform infrared, nuclear magnetic resonance spectroscopy, and transmission electron microscopy. We found that these polyoxotungstates, with 2-4 nm diameters, did not present toxic effects at the tested concentrations. In vitro, POM1 stimulated differentiation of a greater number of dithizone-positive cells (also organized in clusters) than the second nanocompound (POM2). Based on our in vitro studies, we have concluded that both the POMs tested had significant biological activity acting as active stimuli for differentiation of stem cells into insulin-producing cells.

Ultrastructural Analysis of In Vivo Hypoglycemiant Effect of Two Polyoxometalates in Rats with Streptozotocin-Induced Diabetes.

Two polyoxometalates (POMs), synthesized through a self-assembling method, were used in the treatment of streptozotocin (STZ)-induced diabetic rats. One of these nanocompounds [tris(vanadyl)-substituted tungsto-antimonate(III)-anions­POM1] was previously described in the literature, whereas the second [tris-butyltin-21-tungsto-9-antimonate(III)-anions­POM2], was prepared by us based on our original formula. In rats with STZ-induced diabetes treated with POMs (up to a cumulative dose of 4 mg/kg bodyweight at the end of the treatments), statistically significant reduced levels of blood glucose were measured after 3 weeks, as compared with the diabetic control groups (DCGs). Ultrastructural analysis of pancreatic ß-cells (including the mean diameter of secretory vesicles and of their insulin granules) in the treated diabetic rats proved the POMs contribute to limitation of cellular degeneration triggered by STZ, as well as to the presence of increased amounts of insulin-containing vesicles as compared with the DCG. The two POMs also showed hepatoprotective properties when ultrastructural aspects of hepatocytes in the experimental groups of rats were studied. Based on our in vivo studies, we concluded that the two POMs tested achieved hypoglycemiant effects by preventing STZ-triggered apoptosis of pancreatic ß-cells and stimulation of insulin synthesis.

Multistructural biomimetic substrates for controlled cellular differentiation.

Multidimensional scaffolds are considered to be ideal candidates for regenerative medicine and tissue engineering based on their potential to provide an excellent microenvironment and direct the fate of the cultured cells. More recently, the use of stem cells in medicine has opened a new technological opportunity for controlled tissue formation. However, the mechanism through which the substrate directs the differentiation of stem cells is still rather unclear. Data concerning its specific surface chemistry, topology, and its signaling ability need to be further understood and analyzed. In our study, atomic force microscopy was used to study the stiffness, roughness, and topology of the collagen (Coll) and metallized collagen (MC) substrates, proposed as an excellent substrate for regenerative medicine. The importance of signaling molecules was studied by constructing a new hybrid signaling substrate that contains both collagen and laminin extracellular matrix (ECM) proteins. The cellular response-such as attachment capability, proliferation and cardiac and neuronal phenotype expression on the metallized and non-metallized hybrid substrates (collagen + laminin)-was studied using MTT viability assay and immunohistochemistry studies. Our findings indicate that such hybrid materials could play an important role in the regeneration of complex tissues.

Are single nucleotide polymorphisms in the IL-2 gene biomarkers for Hashimoto's thyroiditis?

Background and aims: Hashimoto's thyroiditis (HT) is an autoimmune disorder that can lead to hypothyroidism. The pathophysiology of HT involves the production of antithyroid antibodies that attack the thyroid tissue, causing inflammation and progressive fibrosis. Recent studies demonstrated a strong correlation between Interleukin-2 (IL-2) levels and the development of autoimmune diseases, suggesting that this cytokine may play a crucial role in the pathogenesis of HT. Methods: In this study, we determined the presence of the point mutation +114T/G in the IL-2 gene in patients with HT compared with a control group, and also the serum level of anti-thyroid peroxidase (TPOAbs) and anti-thyroglobulin (TgAbs) antibodies in HT patients with vs. without the mutation. The sequences of the IL-2 gene obtained from subjects were determined by the Sanger sequencing method. Results: Our study did not reveal that the +114T/G polymorphism of the IL-2 gene is a susceptibility or protective factor for HT. No significant correlations were observed between the reference genotype, hetero- and homozygous +114T/G polymorphism and TPOAbs, respectively TgAbs serum levels in HT patients. Conclusions: Further studies of more cases are needed to identify more polymorphisms in the IL-2 gene and study their correlations with HT.

Dracaena arborea alleviates ultra-structural spermatogenic alterations in streptozotocin-induced diabetic rats.

BACKGROUND: Infertility is a common complication in diabetic men and experimental animals, mainly due to loss of germ cells by apoptotic cell death. The aim of this study was to evaluate the effects of aqueous and ethanol extracts of Dracaena arborea in streptozotocin-induced ultra-structural spermatogenic alterations in Wistar rats. METHODS: Diabetic animals were orally treated with Millipore water (10 ml/kg), sildenafil citrate (1.44 mg/kg) or Dracaena arborea aqueous (500 mg/kg) and ethanol (100 mg/kg) extracts for three weeks. A group of non diabetic rats received Millipore water (10 ml/kg) and served as healthy control group. Blood glucose was monitored at the beginning and the end of the study. One day after the last treatment, animals were sacrificed and the testes immediately removed were morphologically observed and prepared for electron microscopy analysis of spermatogenesis. RESULTS: Our results showed that Dracaena arborea was devoid of any anti-hyperglycemic activity. In the untreated diabetic rats, hyperglycemia severely damaged the testes morphology as well as the spermatogenic process as evidenced by the: thickness of basement membrane of the seminiferous tubule; mitochondria alteration; abnormal spermatocyte cells displaying polymorphous nuclei, cytoplasmic vacuolization and necrosis; and disorganization and degeneration of sperm germ cells. Administration of sildenafil citrate and Dracaena arborea extracts to the diabetic rats improved testes morphology and reversed, although not completely, the impairment of spermatogenesis; this alleviating effect was more pronounced in animals treated with the aqueous extract (500 mg/kg) of Dracaena arborea. CONCLUSION: Dracaena arborea improves testes morphology and restores spermatogenesis in type 1 diabetic rats, without having major anti-hyperglycemic properties. These effects could be attributed to saponins, flavonoids, phenols and sterols revealed in this plant, which could be a useful component in the treatment of diabetes-induced testicular dysfunction.

Protective effects of stem bark of Harungana madgascariensis on the red blood cell membrane.

BACKGROUND: Anemia is a condition that has multiple origins. One such origin is the destruction of red blood cells' (RBCs) membrane induced by free radicals. Treatment of anemia could therefore be enhanced by the use of free radicals' scavengers potentially found in some medicinal plants. In this study, the protective effect of Harungana madagascariensis on the RBCs' membrane physiology was investigated in vitro and in vivo. METHODS: In vitro hemolytic anemia was induced by incubation of fresh human RBCs with carbontetrachloride (CCl4) in Olive oil (Oo). Relaxation times of protons excited at 20 MHz (Carr-Purcell-Meiboom-Gill pulse sequence) in the absence or presence of paramagnetic Mn2+ ions (T2i for "extracellular" water and T2a for "intracellular" water, respectively) were determined at several temperatures (25-37°C) via Nuclear Magnetic Resonance (NMR) on a Bruker Minispec spectrometer. Water exchange times (Te) were consequently calculated using the Conlon-Outhred equation: 1/Te = (1/T2a) - (1/T2i). Morphological characteristics (mean cell volume, V, and cell surface area, A) were determined by photonic microscopy and the RBCs' diffusional water permeability (Pd) was calculated as Pd = (1/Te)*(Va/A), where Va is the aqueous volume in the RBC and is about 0.7 of the cell volume (V). The activation energy of the diffusional process (Ea) for the respective temperature range was estimated using the Arrhenius modified equation k = A(T/T0)n*e-Ea/RT. Inhibition of the water diffusion induced by incubation with para-chloro-mercuribenzoic acid (PCMB) at 25, 30 and 37°C was calculated as I(%) = [(Pd control - Pd sample)/Pd control]*100.To investigate the protective influence of the extract on the RBC membrane, inhibition of the water permeability was evaluated on membranes pre-incubated with the Harungana madagascariensis extract. Male rats were used in in vivo investigations. Malondialdehyde (MDA) and cholesterol in the RBC membrane were estimated by induction of lipid peroxidation while the antioxidant properties of catalase (CAT) and superoxide dismutase (SOD) on the membrane were evaluated in regard to their antioxidant properties on the membrane. RESULTS: T2a significantly decreased at each temperature. Te results were higher in both RBCs and RBCs + extract groups incubated with PCMB compared to non-incubated controls, but differences were not statistically significant. A high percentage (73.81 ± 7.22) of RBCs pre-incubated with the extract presented the regular biconcave shape. Inhibition by PCMB of the RBCs' membrane water permeability was increased at 30°C and decreased in the presence of extract (25°C and 37°C), while Ea decreased from 30.52 ± 1.3 KJ/mol to 25.49 ± 1.84 KJ/mol. Presence of the Harungana madagascariensis extract normalized the SOD and CAT activities as well as the MDA and membrane cholesterol concentrations altered by the CCl4-induced oxidative stress. CONCLUSION: Harungana madagascariensis could protect the RBCs' membrane through its antioxidative properties.

Chlamydia trachomatis and the HLA involvement in the development of infection and disease: a narrative review.

Introduction: CT (Chlamydia trachomatis) is among the most common pathogens leading to sexually transmitted diseases. Considering the uncertain mechanism by which HLA polymorphisms influence the CT infection, reinfection, comorbidities or evolution and because there is no consensus regarding the alleles involved in the pathogenesis of the infection, we considered necessary to perform a review to summarize the current knowledge of HLA related to CT. Methods: Pubmed was researched using key terms. Out of the 198 results found, we analyzed articles of all types which describe how the MHC, through HLA alleles, participates in the different stages of CT penetration in the body, including studies about cells or other molecules involved in the process. Results: Almost 40% of the variation in the clinical course of CT infection depends on host genetic factors. There are haplotypes that influence the infection susceptibility/resistance, haplotypes that are involved in the recurrence of the infection, haplotypes that are related to tubal infertility, pelvic inflammatory disease development or trachoma. Antibody to Chsp60 (influenced by MHC genes) has been observed to correlate with late tissue-damaging sequelae. Toll-like receptors were found to increase the susceptibility to CT. The association of HLA-B27 creates susceptibility of reactive arthritis in the organisms infected by CT, but does not influence the carriage of CT. Conclusion: We identified HLA haplotypes belonging both to MHC class l and ll, which influence different stages of CT infection. Genetic risk factors still need research, especially on Caucasians. Studies are moving towards designing a safe and effective vaccine.

Risk factors for suicidal behavior: A Romanian mixed methods study.

OBJECTIVES: This study had two purposes: to explore the main socio-demographic and medical characteristics of the psychiatric patients with a history of suicidal behavior, and to identify the main risk factors underlying the suicidal ideation and acts among psychiatric patients, in the light of two recent theories of suicidal behavior. METHODS: The study is based on a mixed methodological design. During 2019 to 2021, 65 hospitalized psychiatric patients, who committed at least one non-lethal suicide attempt, were investigated using a questionnaire a scales for data collection. Medical records were used to gather data about certain socio-demographic characteristics and the health status of the respondents. Patients also participated in a narrative interview aimed at disclosing their subjective experiences about their past suicidal behavior. RESULTS: The typical psychiatric patient with a history of suicidal behavior, as highlighted by the quantitative analysis, portraits a young old male from an urban environment, childless, educated, having experienced employment problems, being involved in religious activities. The clinical picture of the patient with suicidal antecedents included a moderate or severe level of depression, the presence of socio-emotional loneliness, the manifestation of frequent and long episodes of suicidal ideation, and the intention of committing suicide in order to stop the pain, whose acts resulted in minor injuries. Findings from the qualitative data revealed four major risk factors for the non-lethal suicidal attempts: family disruptions and social problems; economic burdens; a mixture of psychiatric pathologies; and, to a lesser extent, somatic pathologies. CONCLUSION: This study is the first to explore the triggers of suicide acts conducted in a clinical environment in the Balkan region. Implications of the pandemic are also discussed. The findings are useful for designing prevention strategies based on individual psychotherapy and therapeutic or support groups, addressing the main risk factors behind the suicidal ideation and gestures.

Impact of the COVID-19 Pandemic on the Suicidal Behavior in Romania.

Suicide rates reflect the mental health status in certain countries or regions. As the COVID-19 infection developed as a pandemic and governments imposed certain measures to stop the spread of the virus, mental health was affected worldwide. Romania was no exception as the government issued a complete lockdown with restriction regarding travels, social gatherings, transition to working from home and others. The increase in the rate of unemployment, the difficulties in accessing the health services and the social distancing generated by the restrictions had a major impact on lifestyle with a potential surge of psychiatric disorders, with or without history of illness. In this study, we analyzed the changes in regard to the number of suicides and suicide attempts as well as the methods employed and the underlying psychiatric pathology in Cluj County, Romania, throughout the first year of the pandemic. The results had not revealed changes in the overall suicide rate, but in the case of mild cognitive disorders and dementia the suicide figures increased, while a decrease was observed in the number of suicides related to substance use disorders. The long-term effects of the pandemic remain unknown, but there is a clear impact on mental health and measures should be taken in order to prevent suicides.

Determinants of Suicide Ideation in the Romanian Population.

BACKGROUND: Suicide ideation and behaviors are directly linked to the risk of death by suicide. In Romania, as well as worldwide, increased suicide rates were observed in the recent past, more so in the context of the COVID-19 pandemic. The purpose of this study was to investigate the influence of psychosocial factors, quality of life (QOL), and loneliness dimensions and adverse life antecedents on suicide ideation (SI) and prolonged sadness (PS). METHODS: This cross-sectional quantitative research study used a CATI data gathering method to investigate 1102 randomly selected individuals over 18 years of age regarding various determinants of SI and PS. Data were collected in June 2021. Descriptive, inferential, and multivariate statistics were used for data analysis. RESULTS: SI was negatively correlated with all the assessed psychosocial factors, more significantly with family relationships, wealth, health, social relationships, and affective life. Stronger correlations were observed when investigating the state of prolonged sadness, sex, and affective lives along with health and income, which were more influential. SI was negatively correlated with QOL and positively correlated with adverse life events and total loneliness scores. Lesser educated youngsters with reduced overall happiness and a history of depression, self-harm, and trauma were at greater risk of developing SI. CONCLUSIONS: This is the first national study exploring the suicide ideation and prolonged sadness in relation to psychosocial factors, quality of life, and adverse life events. These results have important implications for suicide prevention programs, which should be designed in accordance with similar studies.

Potential Effects of Romanian Propolis Extracts against Pathogen Strains.

The impact of globalization on beekeeping brings new economic, scientific, ecological and social dimensions to this field The present study aimed to evaluate the chemical compositions of eight propolis extracts from Romania, and their antioxidant action and antimicrobial activity against seven species of bacteria, including pathogenic ones: Staphylococcus aureus, Bacillus cereus, Bacillus subtilis, Pseudomonas aeruginosa, Escherichia coli, Listeria monocytogenes and Salmonella enterica serovar Typhimurium. The phenolic compounds, flavonoids and antioxidant activity of propolis extracts were quantified; the presence of flavones and aromatic acids was determined. Quercetin and rutin were identified by HPLC analysis and characterized using molecular descriptors. All propolis samples exhibited antibacterial effects, especially against P. aeruginosa and L. monocytogenes. A two-way analysis of variance was used to evaluate correlations among the diameters of the inhibition zones, the bacteria used and propolis extracts used. Statistical analysis demonstrated that the diameter of the inhibition zone was influenced by the strain type, but no association between the propolis origin and the microbial activity was found.

Cytoarchitectural improvement in Leydig cells of diabetic rats after treatment with aqueous and ethanol extracts of Dracaena arborea (Dracaenaceae).

BACKGROUND AND AIM: Recent studies have demonstrated the androgenic effects of Dracaena arborea in castrated and diabetic rats, but the cytoarchitectural mechanism at the level of Leydig cells (LCs) justifying this improvement in androgens production in diabetic rats has never been examined. We investigated the effects of aqueous and ethanol extracts of D. arborea on diabetes-induced cytoarchitectural impairments of LCs in rats. EXPERIMENTAL PROCEDURE: Besides a normal group, 4 groups of diabetic rats were treated orally with Millipore water (10 ml/kg, diabetic), sildenafil citrate (1.44 mg/kg), aqueous (500 mg/kg) and ethanol (100 mg/kg) extracts of D. arborea for 21 days. On day 22, rats were sacrificed and the testes were removed and prepared for electron microscopic analyses of LCs ultrastructure. RESULTS AND CONCLUSION: The ultrastructure of LCs in control rats was normal, while that in diabetic rats exhibited large heterochromatization in the nuclei, reduced amount of smooth endoplasmic reticulum with no lipid droplets in the cytoplasm, many autophagosomes and degenerated mitochondria containing lots of electron dense granules in the matrix. Interestingly, treatment with D. arborea especially its aqueous extract (500 mg/kg) alleviated these impairments, characterized by a rarification of heterochromatization in the nuclei coupled to an increase and the presence in the cytoplasm of prominent smooth endoplasmic reticulum and a reduction of electron dense granules in the matrix of mitochondria. These alleviating properties of D. arborea on LCs ultrastructure of diabetic rats could explain its androgenic potential. These results are useful for the management of patients suffering from diabetes-induced hypogonadism.

Changes observed in erythrocyte cells exposed to an alternating current.

BACKGROUND AND AIMS: Appliance of electric pulses induces red blood cells (RBCs) membrane poration, membrane aminophospholipid perturbation and alteration of the normal flip-flop process, resulting in various shape changes of the RBCs. We studied morphological and water permeability changes of RBCs bombarded with electrons in an alternating current circuit. METHODS: We used three venous blood samples of 100 mL and an alternating current device. The harvested blood was divided into four experimental sets to be used for various exposure times: 0 hours (control RBCs), 0.5h, 3h and 6h (electric-stimulated RBCs). Following the electric current each of the four sets were further divided into three samples: one for the assessment of the echinocytes/RBCs ratio, another for the electron microscopy study of ultrastructural changes induced by the alternating electrical current and a larger third one for determining water permeability of RCBs by 1H-NMR spectroscopy and morphological measurements. RESULTS: There is a small but statistically significant effect of the RBC exposure to alternating electric current on cell diameters. Exposure to electric current is positively and strongly correlated with the percentage of echinocytes in a duration-dependent manner. There is a strong and statistically significant correlation between electric current exposure and permeability to water as measured by 1H-NMR spectroscopy. CONCLUSION: Following interactions between electric current and RBC membrane, certain modifications were observed in the erythrocyte structure. We attribute the increased cell size to a higher permeability to water and a decreased tonicity. This leads to the transformation of the RBCs into echinocytes.